Novel <sup>89</sup>Zr cell labeling approach for PET-based cell trafficking studies

Aditya Bansal, Mukesh Pandey, Yunus E. Demirhan, Jonathan J. Nesbitt, Ruben J. Crespo-Diaz, Andre Terzic, Atta Behfar, Timothy R DeGrado

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background: With the recent growth of interest in cell-based therapies and radiolabeled cell products, there is a need to develop more robust cell labeling and imaging methods for in vivo tracking of living cells. This study describes evaluation of a novel cell labeling approach with the positron emission tomography (PET) isotope <sup>89</sup>Zr (T<inf>1/2</inf> = 78.4 h). <sup>89</sup>Zr may allow PET imaging measurements for several weeks and take advantage of the high sensitivity of PET imaging. Methods: A novel cell labeling agent, <sup>89</sup>Zr-desferrioxamine-NCS (<sup>89</sup>Zr-DBN), was synthesized. Mouse-derived melanoma cells (mMCs), dendritic cells (mDCs), and human mesenchymal stem cells (hMSCs) were covalently labeled with <sup>89</sup>Zr-DBN via the reaction between the NCS group on <sup>89</sup>Zr-DBN and primary amine groups present on cell surface membrane protein. The stability of the label on the cell was tested by cell efflux studies for 7 days. The effect of labeling on cellular viability was tested by proliferation, trypan blue, and cytotoxicity/apoptosis assays. The stability of label was also studied in in vivo mouse models by serial PET scans and ex vivo biodistribution following intravenous and intramyocardial injection of <sup>89</sup>Zr-labeled hMSCs. For comparison, imaging experiments were performed after intravenous injections of <sup>89</sup>Zr hydrogen phosphate (<sup>89</sup>Zr(HPO<inf>4</inf>)<inf>2</inf>). Results: The labeling agent, <sup>89</sup>Zr-DBN, was prepared in 55% ± 5% decay-corrected radiochemical yield measured by silica gel iTLC. The cell labeling efficiency was 30% to 50% after 30 min labeling depending on cell type. Radioactivity concentrations of labeled cells of up to 0.5 MBq/10<sup>6</sup> cells were achieved without a negative effect on cellular viability. Cell efflux studies showed high stability of the radiolabel out to 7 days. Myocardially delivered <sup>89</sup>Zr-labeled hMSCs showed retention in the myocardium, as well as redistribution to the lung, liver, and bone. Intravenously administered <sup>89</sup>Zr-labeled hMSCs also distributed primarily to the lung, liver, and bone, whereas intravenous <sup>89</sup>Zr(HPO<inf>4</inf>)<inf>2</inf> distributed to the liver and bone with no activity in the lung. Thus, the in vivo stability of the radiolabel on the hMSCs was evidenced. Conclusions: We have developed a robust, general, and biostable <sup>89</sup>Zr-DBN-based cell labeling strategy with promise for wide applications of PET-based non-invasive in vivo cell trafficking.

Original languageEnglish (US)
JournalEJNMMI Research
Volume5
Issue number1
DOIs
StatePublished - Dec 1 2015

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Positron-Emission Tomography
Mesenchymal Stromal Cells
Bone and Bones
Intravenous Injections
Lung
Liver
Membrane Proteins
Cell Tracking
Deferoxamine
Trypan Blue
Silica Gel
Cell- and Tissue-Based Therapy
Isotopes
Dendritic Cells
Radioactivity
Amines
Hydrogen
Melanoma
Myocardium
Phosphates

Keywords

  • Cell labeling
  • In vivo cell tracking
  • Zirconium-89, PET

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

Cite this

Novel <sup>89</sup>Zr cell labeling approach for PET-based cell trafficking studies. / Bansal, Aditya; Pandey, Mukesh; Demirhan, Yunus E.; Nesbitt, Jonathan J.; Crespo-Diaz, Ruben J.; Terzic, Andre; Behfar, Atta; DeGrado, Timothy R.

In: EJNMMI Research, Vol. 5, No. 1, 01.12.2015.

Research output: Contribution to journalArticle

Bansal, Aditya ; Pandey, Mukesh ; Demirhan, Yunus E. ; Nesbitt, Jonathan J. ; Crespo-Diaz, Ruben J. ; Terzic, Andre ; Behfar, Atta ; DeGrado, Timothy R. / Novel <sup>89</sup>Zr cell labeling approach for PET-based cell trafficking studies. In: EJNMMI Research. 2015 ; Vol. 5, No. 1.
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title = "Novel 89Zr cell labeling approach for PET-based cell trafficking studies",
abstract = "Background: With the recent growth of interest in cell-based therapies and radiolabeled cell products, there is a need to develop more robust cell labeling and imaging methods for in vivo tracking of living cells. This study describes evaluation of a novel cell labeling approach with the positron emission tomography (PET) isotope 89Zr (T1/2 = 78.4 h). 89Zr may allow PET imaging measurements for several weeks and take advantage of the high sensitivity of PET imaging. Methods: A novel cell labeling agent, 89Zr-desferrioxamine-NCS (89Zr-DBN), was synthesized. Mouse-derived melanoma cells (mMCs), dendritic cells (mDCs), and human mesenchymal stem cells (hMSCs) were covalently labeled with 89Zr-DBN via the reaction between the NCS group on 89Zr-DBN and primary amine groups present on cell surface membrane protein. The stability of the label on the cell was tested by cell efflux studies for 7 days. The effect of labeling on cellular viability was tested by proliferation, trypan blue, and cytotoxicity/apoptosis assays. The stability of label was also studied in in vivo mouse models by serial PET scans and ex vivo biodistribution following intravenous and intramyocardial injection of 89Zr-labeled hMSCs. For comparison, imaging experiments were performed after intravenous injections of 89Zr hydrogen phosphate (89Zr(HPO4)2). Results: The labeling agent, 89Zr-DBN, was prepared in 55{\%} ± 5{\%} decay-corrected radiochemical yield measured by silica gel iTLC. The cell labeling efficiency was 30{\%} to 50{\%} after 30 min labeling depending on cell type. Radioactivity concentrations of labeled cells of up to 0.5 MBq/106 cells were achieved without a negative effect on cellular viability. Cell efflux studies showed high stability of the radiolabel out to 7 days. Myocardially delivered 89Zr-labeled hMSCs showed retention in the myocardium, as well as redistribution to the lung, liver, and bone. Intravenously administered 89Zr-labeled hMSCs also distributed primarily to the lung, liver, and bone, whereas intravenous 89Zr(HPO4)2 distributed to the liver and bone with no activity in the lung. Thus, the in vivo stability of the radiolabel on the hMSCs was evidenced. Conclusions: We have developed a robust, general, and biostable 89Zr-DBN-based cell labeling strategy with promise for wide applications of PET-based non-invasive in vivo cell trafficking.",
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author = "Aditya Bansal and Mukesh Pandey and Demirhan, {Yunus E.} and Nesbitt, {Jonathan J.} and Crespo-Diaz, {Ruben J.} and Andre Terzic and Atta Behfar and DeGrado, {Timothy R}",
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T1 - Novel 89Zr cell labeling approach for PET-based cell trafficking studies

AU - Bansal, Aditya

AU - Pandey, Mukesh

AU - Demirhan, Yunus E.

AU - Nesbitt, Jonathan J.

AU - Crespo-Diaz, Ruben J.

AU - Terzic, Andre

AU - Behfar, Atta

AU - DeGrado, Timothy R

PY - 2015/12/1

Y1 - 2015/12/1

N2 - Background: With the recent growth of interest in cell-based therapies and radiolabeled cell products, there is a need to develop more robust cell labeling and imaging methods for in vivo tracking of living cells. This study describes evaluation of a novel cell labeling approach with the positron emission tomography (PET) isotope 89Zr (T1/2 = 78.4 h). 89Zr may allow PET imaging measurements for several weeks and take advantage of the high sensitivity of PET imaging. Methods: A novel cell labeling agent, 89Zr-desferrioxamine-NCS (89Zr-DBN), was synthesized. Mouse-derived melanoma cells (mMCs), dendritic cells (mDCs), and human mesenchymal stem cells (hMSCs) were covalently labeled with 89Zr-DBN via the reaction between the NCS group on 89Zr-DBN and primary amine groups present on cell surface membrane protein. The stability of the label on the cell was tested by cell efflux studies for 7 days. The effect of labeling on cellular viability was tested by proliferation, trypan blue, and cytotoxicity/apoptosis assays. The stability of label was also studied in in vivo mouse models by serial PET scans and ex vivo biodistribution following intravenous and intramyocardial injection of 89Zr-labeled hMSCs. For comparison, imaging experiments were performed after intravenous injections of 89Zr hydrogen phosphate (89Zr(HPO4)2). Results: The labeling agent, 89Zr-DBN, was prepared in 55% ± 5% decay-corrected radiochemical yield measured by silica gel iTLC. The cell labeling efficiency was 30% to 50% after 30 min labeling depending on cell type. Radioactivity concentrations of labeled cells of up to 0.5 MBq/106 cells were achieved without a negative effect on cellular viability. Cell efflux studies showed high stability of the radiolabel out to 7 days. Myocardially delivered 89Zr-labeled hMSCs showed retention in the myocardium, as well as redistribution to the lung, liver, and bone. Intravenously administered 89Zr-labeled hMSCs also distributed primarily to the lung, liver, and bone, whereas intravenous 89Zr(HPO4)2 distributed to the liver and bone with no activity in the lung. Thus, the in vivo stability of the radiolabel on the hMSCs was evidenced. Conclusions: We have developed a robust, general, and biostable 89Zr-DBN-based cell labeling strategy with promise for wide applications of PET-based non-invasive in vivo cell trafficking.

AB - Background: With the recent growth of interest in cell-based therapies and radiolabeled cell products, there is a need to develop more robust cell labeling and imaging methods for in vivo tracking of living cells. This study describes evaluation of a novel cell labeling approach with the positron emission tomography (PET) isotope 89Zr (T1/2 = 78.4 h). 89Zr may allow PET imaging measurements for several weeks and take advantage of the high sensitivity of PET imaging. Methods: A novel cell labeling agent, 89Zr-desferrioxamine-NCS (89Zr-DBN), was synthesized. Mouse-derived melanoma cells (mMCs), dendritic cells (mDCs), and human mesenchymal stem cells (hMSCs) were covalently labeled with 89Zr-DBN via the reaction between the NCS group on 89Zr-DBN and primary amine groups present on cell surface membrane protein. The stability of the label on the cell was tested by cell efflux studies for 7 days. The effect of labeling on cellular viability was tested by proliferation, trypan blue, and cytotoxicity/apoptosis assays. The stability of label was also studied in in vivo mouse models by serial PET scans and ex vivo biodistribution following intravenous and intramyocardial injection of 89Zr-labeled hMSCs. For comparison, imaging experiments were performed after intravenous injections of 89Zr hydrogen phosphate (89Zr(HPO4)2). Results: The labeling agent, 89Zr-DBN, was prepared in 55% ± 5% decay-corrected radiochemical yield measured by silica gel iTLC. The cell labeling efficiency was 30% to 50% after 30 min labeling depending on cell type. Radioactivity concentrations of labeled cells of up to 0.5 MBq/106 cells were achieved without a negative effect on cellular viability. Cell efflux studies showed high stability of the radiolabel out to 7 days. Myocardially delivered 89Zr-labeled hMSCs showed retention in the myocardium, as well as redistribution to the lung, liver, and bone. Intravenously administered 89Zr-labeled hMSCs also distributed primarily to the lung, liver, and bone, whereas intravenous 89Zr(HPO4)2 distributed to the liver and bone with no activity in the lung. Thus, the in vivo stability of the radiolabel on the hMSCs was evidenced. Conclusions: We have developed a robust, general, and biostable 89Zr-DBN-based cell labeling strategy with promise for wide applications of PET-based non-invasive in vivo cell trafficking.

KW - Cell labeling

KW - In vivo cell tracking

KW - Zirconium-89, PET

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